Artifical rearrangements of insect ooplasm caused by fixation, and their microkymographic recording

Time-lapse photomicrographs of eggs from the Ichneumonid waspPimpla and from the gall midgeWachtliella reveal that during fixation the ooplasm performs excessive streaming movements when conventional fluids such as Bouin's or Dubosq-Brasil's are used. InWachtliella, intravitelline cleavage may continue for two mitotic cycles during fixation. The cytoplasm of individual egg regions may become displaced up to 45% of the egg's length inPimpla, and up to 14% of the egg's length inWachtliella. Therefore fixed preparations may grossly misrepresent the distribution of egg components in the living state. The changes can be traced back by microkymographic registration of movements during fixation. Microkymograms for this purpose were obtained by projecting a time-lapse film through a slit onto photosensitive material mounted on a rotating drum.The shifting of egg components and the mitotic activity of nuclei fail to occur when the egg is treated with a buffered solution of OsO₄ (1%, 1 min). Subsequent fixation with glutaraldehyde results in satisfactory preservation of microtubules.